GSH Peroxidase Activity Research Articles

Host Prdx6 contributing to the intracellular survival of Brucella suis S2 strain

BackgroundBrucellosis is a worldwide zoonotic infectious disease that is transmitted in various ways and causes great harm to humans and animals. The brucellosis pathogen is Brucella, which mainly resides in macrophage cells and survives and replicates in host cells. However, the mechanisms underlying Brucella survival in macrophage cells have not been thoroughly elucidated to date. Peroxiredoxin 6 (Prdx6) is a bifunctional protein that shows not only GSH peroxidase activity but also phospholipase A2 activity and plays important roles in combating oxidative damage and regulating apoptosis.ResultsRecombinant mouse (Mus musculus) Prdx6 (MmPrdx6) was expressed and purified, and monoclonal antibodies against MmPrdx6 were prepared. Using the Brucella suis S2 strain to infect RAW264.7 murine macrophages, the level of intracellular Prdx6 expression first decreased and later increased following infection. Overexpressing Prdx6 in macrophages resulted in an increase in B. suis S2 strain levels in RAW264.7 cells, while knocking down Prdx6 reduced the S2 levels in cells.ConclusionsHost Prdx6 can increase the intracellular survival of B. suis S2 strain and plays a role in Brucella infection.

THE BENEFIT EFFECT OF GINGER SUPPLEMENTATION AGAINST NICKEL-INDUCED HEPATOTOXICITY IN ALBINO WISTAR RATS

Objective: This study was performed to study the potential capacity effect of ginger on the modulation effects of nickel-induced hepatotoxicity.
 Methods: Thirty-two female albino Wistar rats were divided into four groups of eight each. One served as a control group, the second group (Gi) received ginger 20 g/kg diet, while the third group (Ni) was given nickel 800 mg/L in their drinking water as NiSO46H2O and the fourth group (Ni+Gi) was treated daily with both nickel and ginger. The experiment was lasted for 21 days.
 Results: The exposure to nickel led to a significant decrease in body weight and food intake with an increase of liver weight. Nickel treatment also produced oxidative liver injury characterized by an increase of glucose, cholesterol, triglyceride, total lipids, bilirubin, malondialdehyde (MDA) concentrations and glutamate-pyruvate transaminase, glutamic oxaloacetic transaminase, and alkaline phosphatase activities. Meanwhile, serum total proteins and liver reduced glutathione (GSH) levels, catalase, GSH peroxidase, and GSH superoxide dismutase activities were decreased. These results are substantiated with marked changes in the histopathology, whereas the supplementation of ginger resulted in a restoration of the previous parameters.
 Conclusion: It seems that ginger supplementation is a potent factor for reducing the oxidative severity of nickel hepatotoxicity through its antioxidant action.

Altered skeletal muscle redox status, structural integrity, and ryanodine receptor are associated with contractile dysfunction following systemic iron loading in mice

We previously reported that a moderate degree of systemic iron loading caused muscle functional impairment in mice. Such deterioration was not attributed to oxidative stress since it did not incur protein or lipid oxidative damage. On the other hand, muscle contractile dysfunction was associated with increased thioredoxin protein levels, suggestive of altered redox signaling. Thus, the purpose of this study was to further examine skeletal muscle redox status and potential meditators of contractile dysfunction following a physiologically‐relevant degree of iron loading. Twelve‐week‐old male CD‐1 mice received subcutaneous injections of iron (4 mg iron dextran/200 μL) or vehicle 5 days/week for 2 weeks (n=10/group). One day following the last injection the gastrocnemius muscles were harvested for measurement of biochemical parameters. Systemic iron loading elevated the muscle non‐heme iron (NHI) concentration by approximately 4‐fold (P < 0.001). Although reduced glutathione GSH (P = 0.39) and glutathione peroxidase (GPX) activity (P = 0.82) measures were similar between the two groups, iron‐loaded mice displayed greater glutathione disulfide (GSSG; P = 0.03) and GSSG/GSH ratio (P = 0.03), which are sensitive markers of oxidative stress and redox status. In addition, redox regulator glutaredoxin‐2 (GRX2) protein expression was lower in the muscle of iron‐loaded mice compared with controls (P = 0.04). Muscle structural integrity marker αII‐spectrin at 240 kDa was lower in the iron‐loaded muscles compared with controls (P = 0.02). No differences were observed in its cleavage products at 150 kDa (P = 0.41) or 120 kDa (P = 0.11). Similar to αII‐spectrin, protein levels of focal adhesion α‐actinin were also lower in the iron‐loaded gastrocnemius (P = 0.04). Levels of calstabin‐1 associated with ryanodine receptor (RyR1) were lower in iron‐loaded muscles compared with controls (P = 0.04), while total RyR1 (P = 0.99) was similar between groups. In summary, contractile dysfunction that results from moderate iron loading may be mediated by a disturbance in the muscle redox balance and from changes arising from an increased proteolytic response and aberrant sarcoplasmic reticulum Ca2+ release.Support or Funding InformationSupported by the Syracuse University SOEThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Prenatal exposure to pyrrolizidine alkaloids induced hepatotoxicity and pulmonary injury in fetal rats

Hepatic and pulmonary toxicity in fetal rats induced by pyrrolizidine alkaloids (PAs) was investigated. Retrorsine (RTS) or monocrotaline (MCT) was intragastrically administered during pregnancy. The reduction of body and tail lengths was consistent with body weight loss in PA-exposed fetuses, and pathological lesions in liver and lung were observed only in fetuses. Both PAs reduced fetal serum transaminase activities. The GSH/GSSG ratio, GSH peroxidase and superoxide dismutase activities also decreased but glutathione S-transferase activity increased in fetal lung, especially for MCT. The pyrrole-protein adducts in fetal liver and lung could be detected, and those adducts in RTS fetal lungs were about 65% of those in MCT group. In conclusion, prenatal PAs exposure induced fetal hepatic and pulmonary toxicities through the generation of pyrrole metabolites and oxidative injury. The difference on fetal pulmonary redox homeostasis between two PAs groups might be associated with the content of PAs migrated to fetal lungs.

Glutathione-redox status on hydro alcoholic root bark extract of Premna integrifolia Linn in high fat diet induced atherosclerosis model

BackgroundPremna integrifolia Linn. is a medicinal plant of an Ayurvedic importance and proved to have an anti-inflammatory, anti-diabetic, anti-microbial and hypo-lipidemic activity. Glutathione (GSH) redox status is an important parameter to assess the antioxidant activity of any neutraceuticals. ObjectiveIn order to assess the anti-oxidant potential of hydro alcoholic extract (HAE) of P. integrifolia, this study was aimed to evaluate the GSH redox status in high fat diet induced experimental atherosclerosis. Materials and methodsThe present study comprises sixty Wistar rats and they were divided into six groups: the first group served as control, the second group was fed with high fat diet and the third, fourth and fifth groups were fed with high fat diet along with various concentrations of HAE of 200, 400 and 500 g/kg.b.wt respectively and the sixth group was administered high fat diet along with 10 mg/kg b.wt of atorvastatin for 30 days. GSH-dependent enzymes like GSH-peroxidase (GPx), GSH-reductase (GR) and glucose 6-phosphate dehydrogenase (G6PD) were estimated in hemolysate, kidney, heart and liver of experimental rats. ResultsAnalysis of GSH levels showed a significant decrease in hemolysate, heart and kidney (p < 0.05) and liver (p < 0.01) in high fat-fed rats when compared to control. Activities of GPx, GR and G6PD in hemolysate and heart (p < 0.001), liver and kidney (p < 0.05) in high fat-fed rats when compared to control. Dose-dependent increase was observed in rats treated with various concentrations of HAE. ConclusionThe HAE of root bark of P. integrifolia is proved to have a protective role on antioxidant defense in high fat diet induced atherosclerosis model. As a whole P. integrifolia increases the GSH content in a dose-dependent manner and in turn altered the redox cycle.

Thioredoxin Reductase-1 Inhibition Augments Endogenous Glutathione-Dependent Antioxidant Responses in Experimental Bronchopulmonary Dysplasia.

Background Aurothioglucose- (ATG-) mediated inhibition of thioredoxin reductase-1 (TXNRD1) improves alveolarization in experimental murine bronchopulmonary dysplasia (BPD). Glutathione (GSH) mediates susceptibility to neonatal and adult oxidative lung injury. We have previously shown that ATG attenuates hyperoxic lung injury and enhances glutathione- (GSH-) dependent antioxidant defenses in adult mice. Hypothesis The present studies evaluated the effects of TXNRD1 inhibition on GSH-dependent antioxidant defenses in newborn mice in vivo and lung epithelia in vitro. Methods Newborn mice received intraperitoneal ATG or saline prior to room air or 85% hyperoxia exposure. Glutamate-cysteine ligase (GCL) catalytic (Gclc) and modifier (Gclm) mRNA levels, total GSH levels, total GSH peroxidase (GPx) activity, and Gpx2 expression were determined in lung homogenates. In vitro, murine transformed club cells (mtCCs) were treated with the TXNRD1 inhibitor auranofin (AFN) or vehicle in the presence or absence of the GCL inhibitor buthionine sulfoximine (BSO). Results In vivo, ATG enhanced hyperoxia-induced increases in Gclc mRNA levels, total GSH contents, and GPx activity. In vitro, AFN increased Gclm mRNA levels, intracellular and extracellular GSH levels, and GPx activity. BSO prevented AFN-induced increases in GSH levels. Conclusions Our data are consistent with a model in which TXNRD1 inhibition augments hyperoxia-induced GSH-dependent antioxidant responses in neonatal mice. Discrepancies between in vivo and in vitro results highlight the need for methodologies that permit accurate assessments of the GSH system at the single-cell level.

Impact of a dietary challenge with peroxidized oil on the glutathione redox status and integrity of the small intestine in weaned piglets

Glutathione (GSH) is considered to play an important role in maintaining the integrity of the small intestine. In piglets, altered mucosal GSH levels might therefore be involved in weaning-induced changes of the small intestinal morphology and barrier function. To test this hypothesis, we aimed to challenge the mucosal GSH redox status during the first 28 days after weaning, by feeding diets containing 5% fresh linseed oil (CON), or 2.5% (OF1) or 5% (OF2) peroxidized linseed oil (peroxide value 225 mEq O2/kg oil) and exploring the effects on gut integrity. Piglets were pair-fed and had a total daily feed allowance of 32 g/kg BW. A fourth treatment included animals that were fed the control diet ad libitum (ACON). Animals were sampled at days 5 and 28 post-weaning. The malondialdehyde (MDA) concentration and GSH redox status (GSH/GSSG Eh) were determined in blood, liver and small intestinal mucosa. Histomorphology of the duodenal and jejunal mucosa was determined, and Ussing chambers were used to assess fluorescein isothiocyanate dextran (FD4) and horseradish peroxidase (HRP) fluxes across the mucosa. Results show that peroxidized linseed oil imposed an oxidative challenge at day 28, but not at day 5 post-weaning. At day 28, increasing levels of dietary peroxides to pair-fed pigs linearly increased MDA levels in duodenal and jejunal mucosa. Moreover, FD4 fluxes were significantly increased in OF1 (+75%) and OF2 (+64%) in the duodenum, and HRP fluxes tended (P=0.099) to show similar differences, as compared to CON. This co-occurred with a significant 11 mV increase of the hepatic GSH/GSSG Eh, potentiated by a significantly increased GSH peroxidase activity for treatments OF1 (+47%) and OF2 (+63%) in liver as compared to CON. Furthermore; duodenal HRP flux significantly correlated with the hepatic glutathione disulphide (GSSG) level (r=0.650), as also observed in the jejunum for hepatic GSSG (r=0.627) and GSH/GSSG Eh (r=0.547). The jejunal permeability was not affected, but FD4 and HRP fluxes significantly correlated with the local GSH (r=0.619; r=0.733) and GSSG (r=0.635; r=0586) levels. Small intestinal histomorphology was not affected by dietary lipid peroxides, nor were there any correlations found with the GSH redox system. To conclude, under oxidative stress conditions, jejunal barrier function is related to the local and hepatic GSH redox system. It is suggested that the hepatic GSH system participates in the elimination of luminal peroxides, and thereby impacts on duodenal barrier function.

Effects of constant or intermittent high temperature on egg production, feed intake, and hypothalamic expression of antioxidant and pro-oxidant enzymes genes in laying ducks.

Heat stress is a major environmental factor contributing to lower production of poultry. The objective of present study was to evaluate the influence of constant or intermittent high temperature on the production performance and redox status of plasma and hypothalamus in laying ducks. A total of 288 weight- and laying-matched laying ducks were randomly assigned to 1 of 4 treatments (each with 6 replicates of 12 birds): control, pair-fed, constant high temperature (24 h, 34 ± 1°C), and intermittent high temperature (10 h, 34 ± 1°C). Blood and hypothalamic tissue samples were collected on days 1, 21, and 55 to determine redox status. Average daily feed intake and egg weight was reduced (P < 0.001) during imposition of both high-temperature treatments but was not different (P > 0.05) among the treatments during the recovery period. Lower (P < 0.05) egg mass was observed in pair-fed and intermittent high-temperature treatment during high-temperature period and in constant high temperature during the recovery period. Haugh units from high temperature-treated ducks were significantly lower than those from control or pair-fed ducks (P < 0.05) during the high-temperature period. Both models of heat exposure decreased plasma concentrations of glutathione (GSH) at day 1, and constant high temperature decreased plasma activity of GSH peroxidase (GSH-PX) at day 21 (P < 0.05). Hypothalamic expression of antioxidant genes GSH reductase (GR) and mitochondrial NADH dehydrogenase subunit (Complex Ι) were decreased by both high-temperature treatments at day 1. Hypothalamic expression of genes for pro-oxidant enzymes cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), and cytochrome P450 7A1 (CYP7A1) were decreased (P < 0.05) by both models of high temperature but transcripts of cyclooxygenase-1 (COX-1) of ducks that were pair-fed or were exposed to constant high temperature were increased at day 21. The transcripts of NADPH oxidase 1 (NOX-1) were decreased at day 1 by both high-temperature treatments (P < 0.05) but increased during the recovery period. These results indicate that, for laying ducks, intermittent high temperature caused much greater negative production performance effects than constant high temperature during high-temperature period, but laying ducks exposed to constant high temperature tend to take longer to recover their production performance. High-temperature stress, either constant or intermittent, altered hypothalamic expression of antioxidation and pro-oxidation genes.

BISPHENOL A DOSE- AND TIME-DEPENDENTLY INDUCES OXIDATIVE STRESS IN RAT LIVER MITOCHONDRIA EX VIVO

Objective: The probable toxic effects of bisphenol A (BPA) on different physiological functions have been reported in animal models. The role of BPA in mitochondrial oxidative stress has not been reported till date. The present study is aimed to elucidate dose- and time-dependent oxidative stress generation by BPA, respectively, in rat liver mitochondria in ex vivo model. Methods: The incubation mixture of BPA-treated groups containing mitochondria, 50 mM potassium phosphate buffer (pH 7.4), and different concentrations of BPA (20–160 μM/ml) (dissolved in 12% DMSO) in a final volume of 1.0 ml was incubated at 37°C in incubator for different time durations (30 min–2 h). Whereas, the incubation mixture of control group contained DMSO (12%), mitochondria and 50 mM potassium phosphatebuffer (pH 7.4).’ will be replaced by ‘Whereas, the incubation mixture of control group contained the same constituents except BPA. Result: We have observed significant decrease in mitochondrial intactness incubated with BPA in dose- and time-dependent manner under bright field and confocal microscopic study compared to control. Further, we have observed a decrease in mitochondrial reduced glutathione (GSH) content and increase in lipid peroxidation and protein carbonylation levels in dose- and time-dependent manner in BPA-exposed mitochondria. We have found a significant increase in the activity of Mn-superoxide dismutase and decrease in the activities of GSH peroxidase, GSH reductase, pyruvate dehydrogenase, and other three enzymes of Kreb’s cycle dose and time dependently in BPA-exposed mitochondria. The results indicate that exposure to BPA leads to decrease in intactness of mitochondria and increase in oxidative stress in mitochondria isolated from rat liver in a dose- and time-dependent manner. Conclusion: It can be concluded that the incubation of mitochondria isolated from rat liver with BPA, caused oxidative stress-mediated damages in mitochondria in both dose- and time-dependent manners.

Fumonisin B1 exposure increases Hsp70 expression in the lung and kidney of rats without inducing significant oxidative stress.

The objective of this experiment was to determine whether fumonisin B1 (FB1) added to the diet of rats in a dose of 50 mg/kg changes the production of heat shock protein 70 (Hsp70) in the lungs and kidney of rats. We also studied the effect of this mycotoxin on the antioxidant system of the body. Mature (8 weeks old) male Wistar Crl:WI BR rats (n = 6/group) were fed the toxin-containing diet for 5 days. FB1 resulted in a 7% body weight reduction without significantly changing the feed intake. Western blot analysis of the lungs and kidney demonstrated a substantial (1.4-fold and 1.8-fold, respectively) increase in Hsp70 expression. Alterations could not be detected in the clinical chemical parameters (total protein, albumin, total cholesterol, glucose, creatinine and urea concentrations, and aspartate aminotransferase activity). There was no statistically significant change in malondialdehyde concentrations and the measured antioxidant parameters (the amount of reduced glutathione, GSH and glutathione peroxidase activity, GPx) in the blood plasma, lung and kidney tissue. Thus, it can be concluded that FB1 did not induce oxidative stress in the lungs and kidney, but increased Hsp70 production.

Mice pancreatic islets protection from oxidative stress induced by single-walled carbon nanotubes through naringin.

The growing use of carbon nanotubes (CNTs) emphasizes the importance of its potential toxic effects on the human health. Previous studies proved that CNTs caused oxidative stress and decreased cell viability. On the other hand, reactive oxygen species (ROS) and oxidative stress impaired β-cell functions and reduced the insulin secretion. However, there is not any study on the effects of CNTs on islets and β-cells. Therefore, the present study aimed to evaluate the effects of single-walled CNTs (SWCNTs) on oxidative stress in islets in addition to the protective effects of naringin (NRG) as an antioxidant . We examined the effects of SWCNTs and naringin on islets by 3,4 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay; measurement of insulin secretion, ROS, and malondialdehyde (MDA); activities of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) peroxidase (GSH-Px); and content of GSH and mitochondrial membrane potential (MMP). The MTT assay demonstrated that decreased viability of islets cells was dose-dependent with exposure to SWCNTs. Further studies revealed that SWCNTs decreased insulin secretion and MMP, induced the formation of ROS, increased the level of MDA, and decreased the activities of SOD, GSH-Px, and CAT and content of GSH. Furthermore, the pretreatment of islets with naringin significantly reverted back these changes. These findings revealed that SWCNTs might induce the oxidative stress to pancreatic islets, causing the occurrence of diabetes, and the protective effects of naringin that was mediated by augmentation of the antioxidant defense system of islets. Our research indicated the necessity for further in vivo and in vitro researches on the effects of SWCNTs and naringin on diabetes.

Characterization of the glutathione S-transferases that belong to the GSTFuA class in Ceriporiopsis subvermispora: Implications in intracellular detoxification and metabolism of wood-derived compounds

Glutathione S-transferases (GSTs) of wood-degrading fungi play essential roles in cellular detoxification processes and endogenous metabolism. Fungal GSTs of GSTFuA class are suggested to be involved in lignin degradation. Ceriporiopsis subvermispora is one of the important model fungi of the selective lignin degraders, we found it interesting to study its GSTs. Here, we characterized the activities of two GSTs of the GSTFuA class of C. subvermispora (CsGST63524 and CsGST83044). A high-yield expression systems involving Escherichia coli was developed for each of these enzymes. Both enzymes were found to exhibit GSH-conjugation activity toward 1-chloro-2,4-dinitrobenzene, and GSH-peroxidase activity toward cumene hydroperoxide. Both enzymes showed high GSH-conjugation activity under basic conditions (pH8.0 to 9.0), and the optimum temperature for their activity was 40°C. In addition, three fluorescent compounds were used i.e., methylumbelliferyl acetovanillone was used to monitor etherase activity, and 5-chloromethylfluorescein diacetate and 4-methylumbelliferyl acetate to monitor esterase activity. CsGST83044 exhibited both etherase and esterase activities, while CsGST63524 displayed only esterase activity, which was much higher than that of CsGST83044. These findings imply the functional diversity of the GSTFuA class GSTs of C. subvermispora, suggesting that each protein plays distinctive roles in both the fungal detoxification system and wood compound metabolism.

Hepatoprotective Effect of Essential Oils fromHyptis crenatain Sepsis-Induced Liver Dysfunction

No specific therapeutics are available for the treatment of sepsis-induced liver dysfunction, a clinical complication strongly associated with the high mortality rate of septic patients. This study investigated the effect of the essential oil of Hyptis crenata (EOHc), a lamiaceae plant used to treat liver disturbances in Brazilian folk medicine, on liver function during early sepsis. Sepsis was induced by the cecal ligation and puncture (CLP) model. Rats were divided into four groups: Sham, Sham+EOHc, CLP, and CLP+EOHc. EOHc (300 mg/kg) was orally administered 12 and 24 h after surgery. The animals were sacrificed for blood collection and liver tissue samples 48 h after surgery. Hepatic function was evaluated by measuring serum bilirubin, alkaline phosphatase (ALP), aspartate aminotransferase, and alanine aminotransferase (ALT) levels. The levels of malondialdehyde and the activity of superoxide dismutase, catalase, and GSH peroxidase (GSH-Px) were measured for assessment of oxidative stress. Liver morphology was analyzed by hematoxylin and eosin staining. EOHc normalized serum ALP, ALT, and bilirubin levels and inhibited morphological changes. In addition, we observed that EOHc inhibited elevation in hepatic lipid peroxidation and reduction of the glutathione peroxidase activity induced by sepsis. Our data show that EOHc plays a protective effect against liver injury induced by sepsis.

The effects of oxyresveratrol abrogates inflammation and oxidative stress in rat model of spinal cord injury.

Oxyresveratrol and its glycoside are important natural active materials. As an effective tyrosine kinase inhibitor, oxyresveratrol may prevent herpes virus infection, inflammation and oxidative stress, as well as protect nerves. In addition, it is known to inhibit cell apoptosis following cerebral ischemia. In recent years, oxyresveratrol and its glycoside have been widely investigated, and their useful biological activities have been explored, indicating that they may be worthy of further comprehensive research. The aim of the present study was to evaluate the photoprotective effects of oxyresveratrol and its ability to abrogate inflammation and oxidative stress in a rat model of spinal cord injury (SCI). The authors identified that oxyresveratrol significantly reversed the SCI‑induced inhibition of Basso, Beattie, and Bresnahan scores, inhibited the SCI‑mediated increase in spinal cord water content, significantly suppressed SCI‑induced nuclear factor‑κB/p65, tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6 activities and reversed the malondialdehyde, superoxide dismutase, glutathione (GSH) and GSH peroxidase activities in SCI rats. SCI‑induced granulocyte‑macrophage colony‑stimulating factor (GM‑CSF), inducible nitric oxide synthase (iNOS) and cyclo‑oxygenase‑2 (COX‑2) protein expression was significantly suppressed by oxyresveratrol, and SCI‑mediated inhibition of nuclear factor (erythroid‑derived 2)‑like 2 (Nrf2) protein expression was significantly increased by oxyresveratrol. In conclusion, these results suggest that the effects of oxyresveratrol restores SCI, and abrogates inflammation and oxidative stress in rat model of SCI via the GM‑CSF, iNOS, COX‑2 and Nrf2 signaling pathway.

Effects of glutathione on the survival, growth performance and non-specific immunity of white shrimps (Litopenaeus vannamei)

Reduced glutathione (GSH) is the most abundant non-enzymatic antioxidant present in mammalian cells and the main intracellular defence mechanism against oxidative stress. This study investigated the effects of GSH on survival rate, mean body gain weight, feed efficiency (FE), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, acid phosphatase (ACP), alkaline phosphatase (AKP) activity, GSH peroxidase (GPx) and susceptibility to Vibrio alginolyticus when Pacific white shrimps (Litopenaeus vannamei) were fed with GSH-containing diets. GSH was added to diets at 0.10, 0.20 and 0.30 g/kg during the eight-week breeding experiment. Oral administration of GSH had significantly increased mean body weight gain, FE, PO activity, SOD activity, ACP activity, AKP activity, GPx activity and susceptibility to V. alginolyticus compared with those of the control group (p < .05). Results indicate that GSH exerts both growth-promoting and immunostimulatory effects on Pacific white shrimps (L. vannamei).

42 - Critical Role of Peroxiredoxin 6 in the Repair of Peroxidized Lung Membrane Phospholipids

Peroxidation of membrane lipids represents a severe threat to cellular integrity and its repair is crucial to prevent cell death. Peroxiredoxin 6 (Prdx6), a protein with both GSH peroxidase and phospholipase A2 (PLA2) activities, plays a major role in antioxidant defense of the lung, an organ with very low expression of GPx4.We investigated the role of Prdx6 in the repair of peroxidized cell membranes in intact mouse lungs and isolated pulmonary microvascular endothelial cells (PMVEC) treated with tert-butylhydroperoxide. Lipid peroxidation was evaluated by measurements of: a) thiobarbituric acid reactive substances (TBARS); b) oxidation of diphenyl-1-pyrenylphosphine (DPPP); and c) ferrous- xylenol orange (FOX). The exposure dose in the different models was varied to give a similar increase of about 4-fold over control in lipid peroxidation measurements at end-exposure. Values for lipid peroxidation returned to control levels within 2 h after oxidant removal in wild type (WT) lungs and PMVEC; however,there was essentially no repair in Pxdx6 null lungs or PMVEC. There also was no recovery in lungs and PMVEC that express H26A-Prdx6; this mutation abolishes the phospholipid hydroperoxidase function of Prdx6 but not its ability to reduce H2O2. An intermediate degree of repair was observed with lungs and PMVEC that express either C47S- or D140A-Prdx6; the former mutantion abolishes the peroxidase activity of Prdx6 while the latter abolishes PLA2 activity. Since repair was not seen with Prdx6 knock-out, GPx4 has no role in recovery from peroxidative stress by lung cells while Prdx6 plays a critical role; both the peroxidase and the PLA2 activities of Prdx6 contribute to the recovery process.

N-acetylcysteine in a Double-Blind Randomized Placebo-Controlled Trial: Toward Biomarker-Guided Treatment in Early Psychosis.

Biomarker-guided treatments are needed in psychiatry, and previous data suggest oxidative stress may be a target in schizophrenia. A previous add-on trial with the antioxidant N-acetylcysteine (NAC) led to negative symptom reductions in chronic patients. We aim to study NAC’s impact on symptoms and neurocognition in early psychosis (EP) and to explore whether glutathione (GSH)/redox markers could represent valid biomarkers to guide treatment. In a double-blind, randomized, placebo-controlled trial in 63 EP patients, we assessed the effect of NAC supplementation (2700 mg/day, 6 months) on PANSS, neurocognition, and redox markers (brain GSH [GSHmPFC], blood cells GSH levels [GSHBC], GSH peroxidase activity [GPxBC]). No changes in negative or positive symptoms or functional outcome were observed with NAC, but significant improvements were found in favor of NAC on neurocognition (processing speed). NAC also led to increases of GSHmPFC by 23% (P = .005) and GSHBC by 19% (P = .05). In patients with high-baseline GPxBC compared to low-baseline GPxBC, subgroup explorations revealed a link between changes of positive symptoms and changes of redox status with NAC. In conclusion, NAC supplementation in a limited sample of EP patients did not improve negative symptoms, which were at modest baseline levels. However, NAC led to some neurocognitive improvements and an increase in brain GSH levels, indicating good target engagement. Blood GPx activity, a redox peripheral index associated with brain GSH levels, could help identify a subgroup of patients who improve their positive symptoms with NAC. Thus, future trials with antioxidants in EP should consider biomarker-guided treatment.

Antioxidant and hepatoprotective role of selenium against silver nanoparticles.

Silver nanoparticles (AgNPs) have attracted the most interest in terms of their potential biomedical and industrial applications. However, these nanoparticles have shown their toxic behavior toward environment, living tissues, and organisms. Selenium (Se), an essential trace element, is necessary for various metabolic processes, including protection against oxidative stress and immune function. The present study was undertaken to evaluate the effect of Se against AgNP-induced hepatic oxidative stress. AgNPs were synthesized and then prepared nanoparticles were characterized using various analytical techniques such as UV-visible spectroscopy, X-ray diffraction, and transmission electron microscopy. Rats were administered AgNPs intraperitoneally (5 mg/kg/day) and Se (0.2 mg/kg) was given by gavage. AgNP administration induced hepatic damage as indicated by the serum marker enzymes with reduction in levels of glutathione, and decrease in activities of SOD, CAT, and GSH-peroxidase (P<0.05). Decrease in levels of total antioxidant capacity (TAC) and increase in level of C-reactive protein (CRP) was also observed in AgNP-treated group compared to control group. However, Se markedly attenuated AgNP-induced biochemical alterations, levels of TAC, CRP, and serum transaminases (AST, ALT) (P<0.05). Taken together, these findings suggest that administration of AgNPs produces hepatotoxicity in rats, whereas Se supplementation attenuates these effects.

SEED EXTRACT OF ABELMOSCHUS MOSCHATUS MEDIK REVERSES NAF-INDUCED BEHAVIORAL CHANGES THROUGH NEURODEGENERATION AND OXIDATIVE STRESS IN BRAIN OF RAT

Objective: This study reports protective effect of Abelmoschus moschatus seed extract against sodium fluoride-induced neurodegeneration through oxidative stress, neurohistological, and behavioral observations in Wistar rats.Methods: A total of 20 Wistar rats (around 250 g) were randomly classified into four groups, namely, control, fluoride (NaF), fluoride + A. moschatus seed aqueous extract (AMAE), and fluoride + A. moschatus seed ethanol extract (AMEE). The control group animals received normal tap water, fluoride group received fluoridated water at the rate of 40 mg/kg b. wt., 3rd group rats treated with fluoride (40 mg/kg b. wt.) + AMAE (300 mg/kg b. wt.), and 4th group rats treated with fluoride (40 mg/kg b. wt.) + AMEE (300 mg/kg b. wt.). Neurobehavioral responses of rotarod, hot plate, and maze learning tests and oxidantive stress markers including lipid peroxidation (LPO), GSH levels, superoxide dismutase, CAT, and GSH peroxidase (GPx) activities, and also histology with H and E as well as congo red staining were studied in control, fluoride, and A. moschatus seed extract treated against fluoride groups.Results: Decreased neurobehavioral responses with rotarod, hot plate, and maze and enhanced LPO (p&lt;0.05) levels were found in fluoride received animals. Whereas, the superoxide dismutase (SOD), CAT, GSH, and GPx were decreased (p&lt;0.05) in NaF treatment. The rats received seed extract along with NaF showed significant reversal of behavioral and oxidative stress markers and the effect of ethanol extract was more pronounced than aqueous extract. The fluoride-treated group showed disturbed cell structure and reduced number of cells in H and E as well as congo red staining which was reversed in cell morphology and restored cell number in seed extract against NaF-treated group. As a result of increased LPO, decreased antioxidant system, and decreased number of cells, neurodegeneration was observed resulting in the disturbance in functions associated with reported behavior.Conclusion: Okra with high antioxidants activity, seed extract showed reversal of LPO levels and antioxidant status in the brain tissue. And also plant extract administered rats displayed normal cell structure and number of cells than only fluoride received group. Therefore, the aqueous and ethanolic extract of A. moschatus plant seeds has neuroprotective effects against fluoride-induced motor, nociceptive, learning behavior, and on histological structure of brain through antioxidant mechanism. The ethanol extract has shown more efficacy than aqueous extract.

Peroxiredoxin 6 phospholipid hydroperoxidase activity in the repair of peroxidized cell membranes

Although lipid peroxidation associated with oxidative stress can result in cellular death, sub-lethal lipid peroxidation can gradually resolve with return to the pre-exposure state. We have shown that resolution of lipid peroxidation is greatly delayed in lungs or cells that are null for peroxiredoxin 6 (Prdx6) and that both the phospholipase A2 and the GSH peroxidase activities of Prdx6 are required for a maximal rate of recovery. Like other peroxiredoxins, Prdx6 can reduce H2O2 and short chain hydroperoxides, but in addition can directly reduce phospholipid hydroperoxides. This study evaluated the relative role of these two different peroxidase activities of Prdx6 in the repair of peroxidized cell membranes. The His26 residue in Prdx6 is an important component of the binding site for phospholipids. Thus, we evaluated the lungs from H26A-Prdx6 expressing mice and generated H26A-Prdx6 expressing pulmonary microvascular endothelial cells (PMVEC) by lentiviral infection of Prdx6 null cells to compare with wild type in the repair of lipid peroxidation. Isolated lungs and PMVEC were exposed to tert-butyl hydroperoxide and mice were exposed to hyperoxia (> 95% O2). Assays for lipid peroxidation in wild type control and mutant lungs and cells showed ~4-fold increase at end-exposure. Control lungs and cells showed gradual resolution during a post-exposure recovery period. However, there was no recovery from lipid peroxidation by H26A-Prdx6 lungs or PMVEC. These studies confirm an important role for Prdx6 in recovery from membrane lipid peroxidation and indicate that reduction of H2O2 or short chain hydroperoxides does not play a role in the recovery process.